Description
Key facts
Detection method:Fluorescent,
Sample types:Cell Lysate,
Assay type:Enzyme activity,
Sensitivity:< 1.25 mU/mL,
Assay Platform:Microplate reader
Product details:
Glutathione Peroxidase Activity Assay Kit (Fluorometric) (ab219926) provides a simple method to measure Glutathione Peroxidase activity in cell lysates. This assay is based on the oxidation of glutathione (GSH) to oxidized glutathione (GSSG) catalyzed by Glutathione Peroxidase (GPx). The generated GSSG is recycled to its reduced state GSH by glutathione reductase (GR) and NADPH. The reaction product, NADP
, can be specifically monitored using our newly developed proprietary NADP sensor, which reacts with NADP only to generate a fluorescent product. The signal, which is proportional to the GPx activity present in the sample, can be measured with a fluorescence microplate reader at Ex/Em = 420/480 nm.
This assay can detect activity from as low as 1.25 mU/mL Glutathione Peroxidase in solution. The assay can be performed in a convenient 96-well or 384-well plate format and is easily adapted to automation without a separation or wash step.
Glutathione peroxidase (GPx) is an enzyme family with peroxidase activity to protect the organism from oxidative damage. GPx plays an important role in reducing organic hydroperoxides such as lipid hydroperoxides to their corresponding alcohols, or reducing free hydrogen peroxide to water. It therefore guards against oxidative damage to the cell membranes and other oxidant-sensitive sites in the cell. It has been noticed that altered GPx levels correlate with lesions caused by many comment and complex diseases. GPx level is measured in biological samples as a potential indicator for the potential treatment of cancer, diabetes, neurodegenerative and cardiovascular diseases.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Glutathione Peroxidase (GPx) often referred to as GSH peroxidase is an essential enzyme in the cellular defense system against oxidative damage. It exists as a group of enzymes including the widely studied isoform GPx4. GPx has a molecular weight of approximately 20-23 kDa varying slightly depending on the isoform. This enzyme is localized in many tissues across different organs including the liver kidney and muscles. It catalyzes the reduction of hydrogen peroxide and organic hydroperoxides using glutathione (GSH) as a substrate preventing the formation of free radicals.
Biological function summary
The enzyme plays a significant role in the detoxification of reactive oxygen species. GPx does not usually form a complex with other proteins; instead it acts individually within cells to maintain the balance of oxidative and reductive forces. By converting harmful peroxides into non-toxic alcohols and water it protects cell membranes and other cellular components from oxidative damage. GPx activity is often measured in laboratories using assays to determine its efficiency in catalyzing these reactions.
Pathways
GPx engages in critical cellular antioxidant defense mechanisms. It is an integral part of the glutathione redox cycle where it partners with glutathione reductase another enzyme important for regenerating reduced glutathione. Additionally GPx links to the pentose phosphate pathway providing reduced NADPH necessary for maintaining glutathione in its reduced form. These pathways protect cells from oxidative stress and keep redox homeostasis in check.
Alterations in GPx activity correlate with various health conditions. Researchers link decreased GPx activity to cardiovascular diseases and neurodegenerative disorders highlighting its importance in maintaining cellular health. In cardiovascular diseases the imbalance of oxidative stress pathways involves proteins like superoxide dismutase in addition to GPx worsening the effects of oxidative damage. In neurodegenerative disorders abnormal protein aggregations disturb GPx activities emphasizing the role of antioxidants in disease prevention and management.
